LC3B antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Use primary Ab dilution at 1:200

Protocol tips
- Use primary Ab dilution at 1:200

Publication protocol

Fixed cells were permeabilized with 0.1% Triton X and 1% bovine serum albumin (BSA; Sigma) in PBS, and then blocked with 5% BSA and 5% goat serum (Jackson ImmunoResearch) in PBS for 1 h at room temperature. Cells were stained with primary antibodies diluted in the blocking buffer for 1.5–2 h, and then with fluorescent secondary antibodies (Life Technologies), Hoechst (Life Technologies), and phalloidin (Life Technologies) for 1 h. Images were taken using 20× or 60× objectives on a Nikon A1R (Nikon) or a LSM 710 (Zeiss) confocal microscope. Antibodies against CAV1 (BD Biosciences), clathrin (Cell Signaling Technology), dynamin II (BD Biosciences), EEA1 (BD Biosciences), LAMP1 (BD Biosciences), LC3 (MBL, Novus), CALR (Abcam), GOLGA1 (Golgin97; Molecular Probes) were used.

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Manufacturer protocol

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